Is prnt a pseudogene? Identification of ram Prt in testis and ejaculated spermatozoa.

A hallmark of prion diseases or transmissible spongiform encephalopaties is the conversion of the cellular prion protein (PrP(C)), expressed by the prion gene (prnp), into an abnormally folded isoform (PrP(Sc)) with amyloid-like features that causes scrapie in sheep among other diseases. prnp togeth...

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Autores principales: Jorge Pimenta, Ana Domingos, Pedro Santos, Carla C Marques, Cátia Cantante, Ana Santos, João P Barbas, Maria C Baptista, António E M Horta, Aldino Viegas, Patrícia Mesquita, João Gonçalves, Carlos A Fontes, José A M Prates, Rosa M L N Pereira
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Publicado: Public Library of Science (PLoS) 2012
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spelling oai:doaj.org-article:1474226931504cfa8ebb87062f07acb42021-11-18T07:07:48ZIs prnt a pseudogene? Identification of ram Prt in testis and ejaculated spermatozoa.1932-620310.1371/journal.pone.0042957https://doaj.org/article/1474226931504cfa8ebb87062f07acb42012-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/22937002/?tool=EBIhttps://doaj.org/toc/1932-6203A hallmark of prion diseases or transmissible spongiform encephalopaties is the conversion of the cellular prion protein (PrP(C)), expressed by the prion gene (prnp), into an abnormally folded isoform (PrP(Sc)) with amyloid-like features that causes scrapie in sheep among other diseases. prnp together with prnd (which encodes a prion-like protein designated as Doppel), and prnt (that encodes the prion protein testis specific--Prt) with sprn (shadow of prion protein gene, that encodes Shadoo or Sho) genes, constitute the "prion gene complex". Whereas a role for prnd in the proper functioning of male reproductive system has been confirmed, the function of prnt, a recently discovered prion family gene, comprises a conundrum leading to the assumption that ruminant prnt is a pseudogene with no protein expression. The main objective of the present study was to identify Prt localization in the ram reproductive system and simultaneously to elucidate if ovine prnt gene is transcribed into protein-coding RNA. Moreover, as Prt is a prnp-related protein, the amyloid propensity was also tested for ovine and caprine Prt. Recombinant Prt was used to immunize BALB/c mice, and the anti-Prt polyclonal antibody (APPA) immune response was evaluated by ELISA and Western Blot. When tested by indirect immunofluorescence, APPA showed high avidity to the ram sperm head apical ridge subdomain, before and after induced capacitation, but did not show the same behavior against goat spermatozoa, suggesting high antibody specificity against ovine-Prt. Prt was also found in the testis when assayed by immunohistochemistry during ram spermatogenesis, where spermatogonia, spermatocytes, spermatids and spermatozoa, stained positive. These observations strongly suggest ovine prnt to be a translated protein-coding gene, pointing to a role for Prt protein in the ram reproductive physiology. Besides, caprine Prt appears to exhibit a higher amyloid propensity than ovine Prt, mostly associated with its phenylalanine residue.Jorge PimentaAna DomingosPedro SantosCarla C MarquesCátia CantanteAna SantosJoão P BarbasMaria C BaptistaAntónio E M HortaAldino ViegasPatrícia MesquitaJoão GonçalvesCarlos A FontesJosé A M PratesRosa M L N PereiraRosa M L N PereiraPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 7, Iss 8, p e42957 (2012)
institution DOAJ
collection DOAJ
language EN
topic Medicine
R
Science
Q
spellingShingle Medicine
R
Science
Q
Jorge Pimenta
Ana Domingos
Pedro Santos
Carla C Marques
Cátia Cantante
Ana Santos
João P Barbas
Maria C Baptista
António E M Horta
Aldino Viegas
Patrícia Mesquita
João Gonçalves
Carlos A Fontes
José A M Prates
Rosa M L N Pereira
Rosa M L N Pereira
Is prnt a pseudogene? Identification of ram Prt in testis and ejaculated spermatozoa.
description A hallmark of prion diseases or transmissible spongiform encephalopaties is the conversion of the cellular prion protein (PrP(C)), expressed by the prion gene (prnp), into an abnormally folded isoform (PrP(Sc)) with amyloid-like features that causes scrapie in sheep among other diseases. prnp together with prnd (which encodes a prion-like protein designated as Doppel), and prnt (that encodes the prion protein testis specific--Prt) with sprn (shadow of prion protein gene, that encodes Shadoo or Sho) genes, constitute the "prion gene complex". Whereas a role for prnd in the proper functioning of male reproductive system has been confirmed, the function of prnt, a recently discovered prion family gene, comprises a conundrum leading to the assumption that ruminant prnt is a pseudogene with no protein expression. The main objective of the present study was to identify Prt localization in the ram reproductive system and simultaneously to elucidate if ovine prnt gene is transcribed into protein-coding RNA. Moreover, as Prt is a prnp-related protein, the amyloid propensity was also tested for ovine and caprine Prt. Recombinant Prt was used to immunize BALB/c mice, and the anti-Prt polyclonal antibody (APPA) immune response was evaluated by ELISA and Western Blot. When tested by indirect immunofluorescence, APPA showed high avidity to the ram sperm head apical ridge subdomain, before and after induced capacitation, but did not show the same behavior against goat spermatozoa, suggesting high antibody specificity against ovine-Prt. Prt was also found in the testis when assayed by immunohistochemistry during ram spermatogenesis, where spermatogonia, spermatocytes, spermatids and spermatozoa, stained positive. These observations strongly suggest ovine prnt to be a translated protein-coding gene, pointing to a role for Prt protein in the ram reproductive physiology. Besides, caprine Prt appears to exhibit a higher amyloid propensity than ovine Prt, mostly associated with its phenylalanine residue.
format article
author Jorge Pimenta
Ana Domingos
Pedro Santos
Carla C Marques
Cátia Cantante
Ana Santos
João P Barbas
Maria C Baptista
António E M Horta
Aldino Viegas
Patrícia Mesquita
João Gonçalves
Carlos A Fontes
José A M Prates
Rosa M L N Pereira
Rosa M L N Pereira
author_facet Jorge Pimenta
Ana Domingos
Pedro Santos
Carla C Marques
Cátia Cantante
Ana Santos
João P Barbas
Maria C Baptista
António E M Horta
Aldino Viegas
Patrícia Mesquita
João Gonçalves
Carlos A Fontes
José A M Prates
Rosa M L N Pereira
Rosa M L N Pereira
author_sort Jorge Pimenta
title Is prnt a pseudogene? Identification of ram Prt in testis and ejaculated spermatozoa.
title_short Is prnt a pseudogene? Identification of ram Prt in testis and ejaculated spermatozoa.
title_full Is prnt a pseudogene? Identification of ram Prt in testis and ejaculated spermatozoa.
title_fullStr Is prnt a pseudogene? Identification of ram Prt in testis and ejaculated spermatozoa.
title_full_unstemmed Is prnt a pseudogene? Identification of ram Prt in testis and ejaculated spermatozoa.
title_sort is prnt a pseudogene? identification of ram prt in testis and ejaculated spermatozoa.
publisher Public Library of Science (PLoS)
publishDate 2012
url https://doaj.org/article/1474226931504cfa8ebb87062f07acb4
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